CN108467869A - The application of soybean sucrose transporter important gene GmSUT6 - Google Patents
The application of soybean sucrose transporter important gene GmSUT6 Download PDFInfo
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Abstract
The present invention discloses a kind of application of soybean sucrose transporter important gene GmSUT6, is related to plant genetic engineering and biotechnology.The present invention identifies the sucrose transporter gene GmSUT6 of induced expression in a low-phosphorous root nodule, the expression of the gene is by low-phosphorous regulation and control by real time fluorescence quantifying PCR method in soybean.Under the conditions of low-phosphorous, overexpression GmSUT6 has the function of Nodule Growth development in Transgenic soybean plants, there is significant impact to the symbiosis of soybean and rhizobium, this is to illustrating biological function of the SUT genes in legume and rhizobium symbiosis, to regulate and control the beneficial symbiosis important in inhibiting between plant and root nodule.
Description
Technical field
The present invention relates to plant genetic engineerings and biotechnology, and in particular to a kind of soybean sucrose transporter is important
The application of gene GmSUT6.
Background technology
Higher plant is the autotroph being made of autotrophic organ and heterotrophism organ.Heterotrophism organ needs the light of autotrophic organ
Product is closed to maintain growth and development.The photoassimilates 80% synthesized in plant maturation blade etc. " source " organ all pass through plant
Skeleton transports root, flower, fruit etc. " library " organ.Sucrose is one of green plants photoassimilates, and most
Photosynthate trasport and the principal mode of distribution in higher plant body (Yang Shuan etc., 2005；Bai Xuemei etc., 2006).Sucrose is being planted
The mode of orientation transport and distribution not only regulates and controls the entire growth and development process of plant in strain, also determines the yield and product of crop
Matter.And durings " source " organ of sucrose loading, the long-distance transportation of bast and the unloading of " library " organ etc., sucrose transports egg
In vain (Sucrose Transporter or Sucrose Carrier, SUT or SUC) play an important role (Williams et
al.,2000；Wu Xiaodan etc., 2006；Ruan Yanye etc., 2007).Sucrose transporter belongs to easyization diffusion superfamily (Major
Facilitator Superfamily, MFS), there is typical 12 transmembrane domains (Stolz et al., 1999).Sucrose
Transport protein not only plays a crucial role in the loading of bast and unloading, but also is distributed in plant and homobium or pathogen carbon
In it is same it is most important (Wahl et al., 2010；Wippel et al.,2010).
Rhizobium (Rhizobium, R) are the nonspore-bearing agrobacteriums of a kind of Gram-negative, are distributed in the soil very extensively,
Mutually beneficial symbiosis can be formed with legume and some non-leguminous plants.Rhizobium are by infecting host plant
Root system form a kind of special structure --- root nodule.Being formed by root nodule can help host plant by the free state in air
Nitrogen is converted into the ammonium nitrogen that can be directly absorbed and utilized, and as return, host plant can provide photosynthetic carbon for rhizobium, normal for it
Growth and development, this phenomenon is referred to as symbiotic nitrogen fixation (Qin et al., 2011).Therefore, total in host plant and rhizobium
In raw system, sucrose transporter necessarily plays vital effect.
Although in arabidopsis and clover, sucrose transporter family (SUTs) has correlative study, in soybean
Still there is not been reported, and concrete function of the gene family member in root nodule cogeneration system is not known yet.
Invention content
In order to overcome the disadvantages and deficiencies of the prior art, the purpose of the present invention is to provide a kind of soybean sucrose transporters
The application of important gene GmSUT6.
The purpose of the invention is achieved by the following technical solution：
A kind of soybean sucrose transporter important gene GmSUT6 of present invention offer is in low-phosphorous root nodule in terms of induced expression
Application.
The nucleotides sequence of the soybean sucrose transporter important gene GmSUT6 is classified as Glyma.10G217900 institutes
Show, the amino acid sequence of the protein of GmSUT6 gene codes is shown in Glyma.10G217900.
The present invention identifies induced expression in a low-phosphorous root nodule by real time fluorescence quantifying PCR method in soybean
Sucrose transporter gene GmSUT6, the expression of the gene is by low-phosphorous regulation and control.Under the conditions of low-phosphorous, excess in Transgenic soybean plants
Expression GmSUT6 has the function of Nodule Growth development, and the beneficial symbiosis between regulation and control plant and root nodule has important meaning
Justice.
The present invention also provides a kind of applications of soybean sucrose transporter important gene GmSUT6 in plant breeding；Into one
Step, the application in cultivating genetically modified plants.
The plant is dicotyledonous legume.
The dicotyledonous legume is soybean.
Gene GmSUT6 and protein provided by the invention can regulate and control the growth of root nodule in the transgenosis root system comprising it
Development.
The primer pair for expanding above-mentioned GmSUT6 full length genes or its any segment belongs to the scope of protection of the present invention.
The present invention also provides the recombinant expression carriers containing above-mentioned GmSUT6 genes, can use existing plant expression vector structure
Build the recombinant expression carrier containing GmSUT6 genes.The plant expression vector includes double base agrobacterium vector etc., such as pYLRNAi
(Hu Xuxia and Liu Yaoguang, 2006, Molecular Plant Breeding) or other derivative plant expression vectors.
The present invention also provides a kind of genetic engineering bacteriums, contain above-mentioned recombinant expression carrier.
The invention further relates to cells, and it includes the GmSUT6 genes or recombinant expression carrier of the present invention.The cell can be with
It is plant cell, such as legume cell or microbial cell, such as bacterium or fungal cell, such as yeast cells.Institute
It can be separation, in vitro, part culture or that be plant to state cell.
The invention further relates to plant or plant parts, vegetable material, and vegetable seeds, it includes the cells of the present invention.Institute
It can be legume, such as Kidney bean and soybean to state plant, can also be other plants, such as monocotyledon such as rice, small
Wheat, barley, corn, sorghum, sugarcane, oat, rye etc. or other dicotyledons for example tobacco, sunflower, beet, capsicum,
Potato, tomato etc..Further relate to the transgenic seed from the plant.
The invention further relates to the method for production plant, this method includes：It is planted from the Plant cell regeneration transgenosis of the present invention
Object, or by plant and another plant hybridization of the present invention.
The invention further relates to the plants of the method for present invention production.
The present invention also provides a kind of soybean sucrose transporter important gene GmSUT6 in regulation and control plant and rhizobium symbiosis
Application in relationship；Further, regulation and control plant and the application in rhizobium symbiosis preparation are being prepared.
The plant is dicotyledonous legume.
The dicotyledonous legume is soybean.
The invention further relates to the GmSUT6 genes of the present invention or recombinant vector in regulation and control plant and rhizobium symbiosis shape
Purposes in, including prepare transgenosis plant and the preparation for preparing regulation and control plant and rhizobium formation symbiosis.
The invention further relates to the method that regulation and control plant forms symbiosis with rhizobium, this method includes preparing to contain this hair
The plant of bright GmSUT6 genes or recombinant vector.For example, the method may include turn from the Plant cell regeneration of the present invention
Gene plant or by plant and another plant hybridization of the present invention.
A preferred embodiment provided by the present invention is that said gene GmSUT6 is imported soybean hypocotyl injection to lure
In the root system led, transgenic line is obtained；The growth of the transgenic line, including biomass and Nodule Growth happen it is bright
Aobvious variation.
The gene GmSUT6 can import receptor soybean hypocotyl root system by the recombinant expression carrier.
Carrying the plant expression vector of the gene GmSUT6 of the present invention can turn for example, by Agrobacterium-medialed transformation method
Change into soybean hypocotyl root system.
The present invention has the following advantages and effects with respect to the prior art：
Though the SUT gene families belonging to gene GmSUT6 have been cloned and have reported in arabidopsis and clover etc., it is in beans
Biological function in terms of section crop and root nodule symbiosis is not known.The gene GmSUT6 of the present invention is total to soybean and rhizobium
Raw to have significant impact, this is to illustrating biological function of the SUT genes in legume and rhizobium symbiosis, to regulate and control
Beneficial symbiosis important in inhibiting between plant and root nodule.
Description of the drawings
Fig. 1 is expression pattern analysis of the GmSUTs gene families in root nodule；Plant uses solution culture, in 5 μM of P
(LP) or under the conditions of 500 μM of P (HP), Rhizobium Inoculation harvests after handling 40 days；Data are the flat of biology repetition three times in figure
Mean value and standard error, * indicate same gene significant difference (P between high low-phosphorous processing<0.05).
Fig. 2 is the sugared absorption function verification analysis of GmSUT6；Wherein SUSY7/ura3 compares for sucrose defective yeast, CK
To be transferred to the control of pDR196 zero loads, AtSUC2 is the positive control for being transferred to arabidopsis sucrose transporter AtSUC2 genes,
GmSUT6 is the bacterial strain for being transferred to soybean sucrose transporter GmSUT6.
Fig. 3 is the expressive site analysis of GmSUT6 promoters driving GUS.
Fig. 4 is the Subcellular Localization of GmSUT6；First row is the Tobacco Epidermis for converting empty carrier in figure, immediately
It is the tobacco cell for converting GFP-GmSUT6 carriers, and figure is observation shooting GFP and film positioning under Laser Scanning Confocal Microscope
The fluorescent co-location of Marker mCherry.
Fig. 5 is the expression quantity detection of overexpression GmSUT6 transfer-gen plants；Wherein, CK expressions are transferred to pYLRNAi.2 skies
The control of load, OX indicate overexpression GmSUT6 transfer-gen plants.Data are the average value and mark that five secondary pollutants repeat in figure
Standard is accidentally.* it indicates same and meets under bacterium processing significant difference (P between different transgenic lines<0.05).
Fig. 6 is shadows of the overexpression GmSUT6 to Soybean transgenic plant dross situation (A) and plant strain growth situation (B)
It rings；CK indicates that the control for being transferred to pYLRNAi.2 zero loads, OX indicate overexpression GmSUT6 transfer-gen plants.Data are five in figure
The average value and standard error that secondary pollutant repeats.* it indicates same and meets under bacterium processing significant difference (P between different transgenic lines<
0.05)。
Specific implementation mode
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited
In this.
Test method used in following embodiments is conventional method unless otherwise specified；Used material, examination
Agent etc., unless otherwise specified, for the reagent and material commercially obtained.
Embodiment 1
Expression pattern analysis of the GmSUTs gene families in root nodule：In the soybean genome database announced, lead to
Homologous comparison is crossed, soybean sucrose transporter gene is predicted and shares 11 members, then by quantitative PCR technique, is determined
Expression of the GmSUTs family members gene in high low-phosphorous processing root nodule.
Using 1 soybean varieties, 2 nutrient water are arranged with the training method of water planting in phosphorus efficiency Guangdong spring 04-5 (YC04-5)
Flat, respectively 5 μM of P (LP) and 500 μM of P (HP), each processing are arranged 3 biology and repeat, Rhizobium Inoculation BXYD3 40
It is harvested after it.Root nodule RNA is extracted, reverse transcription further uses the expression of quantitative PCR detection GmSUTs family gene members at cDNA
Pattern.The house-keeping gene EF-1a of soybean is as internal reference.Primer for quantitative PCR detection gene expression amount is respectively：
The primer of soybean EF-1a genes is：
EF-1a F：5′-TGAACCACCCTGGTCAGATT-3′(SEQ ID NO:1)
EF-1a R：5′-TCCAGCATCACCATTCTTCA-3′(SEQ ID NO:2)
The primer of GmSUTs genes is：
GmSUT1F：5′-GTTCCTCTTGCGATTACTTACAG-3′(SEQ ID NO:3)
GmSUT1R：5′-CACCACCAAACCATTTATCCC-3′(SEQ ID NO:4)
GmSUT2F：5′-CAGGACAAGGTTTATCTTTGGGAG-3′(SEQ ID NO:5)
GmSUT2R：5′-ACCACCAAACAAGGAATCCC-3′(SEQ ID NO:6)
GmSUT3F：5′-TGGTCCCACAGATAATAGTGTC-3′(SEQ ID NO:7)
GmSUT3R：5′-GCCAACACAGCTATGAGTCC-3′(SEQ ID NO:8)
GmSUT4F：5′-CAAATATCCTAATGGCTGTCTGCT-3′(SEQ ID NO:9)
GmSUT4R：5′-CGCTGTAAGTGATTGCAAGTG-3′(SEQ ID NO:10)
GmSUT5F：5′-CTCTCACTTGGTTGTCTTGG-3′(SEQ ID NO:11)
GmSUT5R：5′-AAATGCACCTTCTCTAACTCC-3′(SEQ ID NO:12)
GmSUT6F：5′-CTAAGATGGCCCAACATTCTC-3′(SEQ ID NO:13)
GmSUT6R：5′-GCTGTAAGTAATCGCGAGTG-3′(SEQ ID NO:14)
GmSUT7F：5′-CTCTTGCTATTACTTTCAGCGT-3′(SEQ ID NO:15)
GmSUT7R：5′-CACCACCAAACCACTTATCC-3′(SEQ ID NO:16)
GmSUT8F：5′-CTCTCACAACAATGGAGGAGC-3′(SEQ ID NO:17)
GmSUT8R：5′-ACATAGCCAAATAAATGAGGCCC-3′(SEQ ID NO:18)
GmSUT9F：5′-CAAGGTCTATCTTTGGGTGTC-3′(SEQ ID NO:19)
GmSUT9R：5′-GAAAGTGAAACATGTAAAGCCC-3′(SEQ ID NO:20)
GmSUT10F：5′-TTCCTCTTGCGATTACTTTCAG-3′(SEQ ID NO:21)
GmSUT10R：5′-AAACAAAGCATCCCAAGGAC-3′(SEQ ID NO:22)
GmSUT11F：5′-TCCAATAGTACGCAAACTGAG-3′(SEQ ID NO:23)
GmSUT11R：5′-CCATGACAACCAAGTGAGAG-3′(SEQ ID NO:24)
Quantitative PCR response procedures and condition are：RNA sample is inverted into gained cDNA and dilutes 20 times as quantitative PCR reaction
Template.It chooses appropriate cDNA stostes and does the template that gradient dilution is standard curve.20 μ L reaction systems are used in experiment, including：
The PowerUP SYBR Green Master Mix of 10 μ L, 10 μM of forward and reverse primers of each 0.6 μ L, the 2 diluted cDNA of μ L, most
Afterwards with sterile ddH2O is mended to 20 μ L.Reaction condition is 95 DEG C and is denaturalized 1 minute, is then denaturalized 15 seconds for 95 DEG C, 60 DEG C are annealed and prolonged
It stretches 60 seconds, carries out 40 cycles.With the StepOne of Applied BiosystemsTMSoftware v2.2.2 calculate each sample
The expression quantity of product.The results are shown in Figure 1 (unlisted gene is not expressed in root nodule in figure).
The result shows that：In GmSUTs family members, GmSUT6 is in low-phosphorous root nodule by induced strong up-regulated expression.
Embodiment 2
1, the clone of GmSUT6 genes：Using soybean YC03-3 leaf portions cDNA as masterplate, with upstream specific primer 5 '-
atcgcccgggCCTGCTGCTACAATATGGAG-3′(SEQ ID NO:And downstream special primer 5 ' -25)
atcgctcgagCACGGTCAACTCTGATTAGTG-3′(SEQ ID NO:26) the ORF full length sequences of GmSUT6 genes are expanded
1566bp, and comparison is sequenced, it obtains GmSUT6 coded sequences and sees that Glyma.10G217900, corresponding protein sequence are shown in
Glyma.10G217900。
2, the structure of yeast covering experimental vehicle：Using soybean YC03-3 leaf portions cDNA as masterplate, with upstream specific primer 5 '-
atcgcccgggCCTGCTGCTACAATATGGAG-3′(SEQ ID NO:And downstream special primer 5 ' -25)
atcgctcgagCACGGTCAACTCTGATTAGTG-3′(SEQ ID NO:26) the ORF full length sequences of GmSUT6 genes are expanded
1566bp, PCR recycle segment, after carrying out double digestion to segment and purpose carrier by Sma I and Xho I, by GmSUT6 genes
It is connected to purpose carrier pDR196.Escherichia coli DH10B is converted, errorless rear conversion sucrose defective yeast bacterial strain SUSY7/ is sequenced
Ura3, for carrying out yeast covering experiment.Experimental result is shown in Fig. 2.The result shows that：GmSUT6 has the function of transporting sucrose.
Embodiment 3
GmSUT6 gene promoters are cloned and vector construction：The structure of promoter Analysis expression vector：Conventionally,
The leaves genomic DNA of extraction soybean YC03-3 genotype uses upstream specific primer using soybean leaf portion genomic DNA as template
5′-atcggaattcAGCGTATAAGCATACTAAAC-3′(SEQ ID NO:And downstream special primer 5 ' -27)
atcgtctagaATTGTAGCAGCAGGAAGAAAG-3′(SEQ ID NO:28) GmSUT6 promoter 2547bp segments are expanded,
After PCR fragment recycling, after carrying out double digestion to recycling segment and purpose carrier by EcoR I and Xba I, GmSUT6 is started
Subbase is because being connected to purpose carrier pTF102.Escherichia coli DH10B is converted, errorless rear conversion Agrobacterium tumefaciems EHA101 is sequenced,
For the whole strain conversion of Agrobacterium tumefaciens mediated soybean cotyledon node (Wang et al., 2009).
The detection of the whole strain transformation plant of promoter：It is chosen on transfer-gen plant in the trifoliolate leaf being just fully deployed
A piece of, half is marked with marking pen, the other half dipped with cotton swab diluted herbicide (Originate from France) it applies
It is put on the front of blade, observation blade changes after 2~3d.It is withered or have yellow spotting production if blade occurs chlorosis, turns yellow
It is raw then illustrate the plant not antiweed, for negative non-transgenic material；If blade does not change, illustrate its plant
It may be positive plant inside to have Herbicid resistant.Meanwhile carrying out the qualitative detection of gus protein expression activity.Acquire fresh leaf
Or root is placed in culture dish, the GUS dye liquors prepared in right amount, which are added, makes sample to be tested be dipped into completely in GUS dye liquors, is put into 37 DEG C
In insulating box when obvious blue occurs in sample, dye liquor is outwelled.After impregnating 5min rinsing samples with 50% ethyl alcohol, it is added 70%
Ethyl alcohol impregnates until observing GUS staining conditions after sample decoloration completely.
The whole strain transformation plant of promoter is selected using sand culture test for GmSUT6 tissue expression positioning analysis
ProGmSUT6-35 is BXYD3, low 500 μM of Ca (NO of nitrogen (LN) for examination rhizobium3)2·4H2O.The K of low nitrogen processing+Use K2SO4
Polishing.Each processing is arranged 4 biology and repeats.Routine Management, florescence collect new fresh quench and root nodule sample, clean, and carry out
GUS vital stainings.As a result see Fig. 3.The result shows that：GmSUT6 is primarily located in root system in root nodule.
Embodiment 4
The structure of subcellular localization experimental vehicle：Using soybean YC03-3 leaf portions cDNA as masterplate, with upstream specific primer 5 '-
GGGGacaagtttgtacaaaaaagcaggcttcATGGAGCCTCTCTCTTCCAC-3′(S EQ ID NO:29) and downstream is special
Different primer 5 '-GGGGaccactttgtacaagaaagctgggtcCACGGTCAACTCTGATTAGTG-3 ' (SEQ ID NO:30)
ORF full length sequences 1566bp, PCR the recycling segment for expanding GmSUT6 genes, is connected GmSUT6 genes by Gatway technologies
To purpose carrier pMDC43.Escherichia coli DH10B is converted, errorless rear conversion Agrobacterium GV3101 is sequenced, for carrying out subcellular
Positioning experiment.
For the Subcellular Localization of GmSUT6, using the method for Tobacco Epidermis instantaneous conversion.First fusion is had
The GV3101 bacterium solutions of GmSUT6 genes and the GV3101 bacterium solutions for being transferred to film Marker plasmids 1008 are shaken bacterium and are stayed overnight, and are then centrifuged for,
With including 10mM MES, 10mM MgCl2With the re-suspension liquid resuspended bacterium solution of 150 μM of AS.22~24 DEG C of 3~4h of light culture, will
The two after mixing, is converted into the tobacco leaf of 3~4 week old in equal volume, after converting 3 days, at laser co-focusing 488nm (GFP)
With observation fluorescence at 587nm (mCherry).Experimental result is shown in Fig. 4.The result shows that：GmSUT6 is located on plasma membrane.
Embodiment 5
1, the structure of overexpression GmSUT6 carriers：Using the leaf portion cDNA of soybean YC03-3 genotype as template, upstream is used
5 '-atcg of special primeragatctgaCCTGCTGCTACAATATGGAG-3′(SEQ ID NO:And downstream special primer 5 ' -31)
atcgacgcgtCACGGTCAACTCTGATTAGTG-3′(SEQ ID NO:32) the ORF full length sequences of GmSUT6 are expanded
After 1566bp, PCR recycle segment, after carrying out double digestion to segment and purpose carrier by Bgl II and Mlu I, by GmSUT6 bases
Because being connected to purpose carrier pYLRNAi.2.Escherichia coli DH10B is converted, errorless rear transforming agrobacterium rhizogenes K599 is sequenced, is used for
The soybean hypocotyl injection conversion that agrobacterium rhizogenes mediates.
2, the acquisition of Soybean transgenic plant：The overexpression GmSUT6 expression vector plasmids built are converted to root of hair
In Agrobacterium, the soybean hypocotyl injection conversion that is mediated using agrobacterium rhizogenes obtain transgenosis chimeric plant (Li et al.,
2014), subsequent phenotypic evaluation uses transgenosis to be fitted into strain.
3, the detection of Soybean transgenic plant
Overexpression GmSUT6 hypocotyls inject the detection of converting material：Clip part single plant hypocotyl wound is grown
Adventitious root extracts RNA and is reversed to cDNA as template, and Plasmid DNA is as positive control, the root system cDNA of Non-transgenic soybean
As negative control, with upstream specific primer 5 '-GCTGTTATGCGGCCATTGTC-3 ' (SEQ ID NO:33) and downstream is special
Primer 5 '-GACGTCTGTCGAGAAGTTTC-3 ' (SEQ ID NO:34) amplification hygromycin gene segment carries out PCR inspections
It surveys.In addition, the effect of quantitative PCR detection overexpression is further used, it is as described above with soybean house-keeping gene in quantitative PCR assays
EF-1a is reference gene (primer sequence：SEQ ID NO:1,2), relative expression quantity is purpose gene GmSUT6 (primer sequences：
SEQ ID NO:13,14) ratio of expression quantity and house-keeping gene expression quantity.Confirm had by PCR detections and quantitative PCR
The different transgenic lines of effect.As a result see Fig. 5.
4, influences of the overexpression GmSUT6 to Soybean transgenic plant dross situation：Harvest Rhizobium Inoculation 6 weeks not
With transgenic line and dross situation is measured, including：Root nodule number and root nodule fresh weight etc..
Fig. 6 A are influences of the overexpression GmSUT6 to Soybean transgenic plant dross situation.Wherein, plant uses sand
Training experiment, sand ratio：Native 600g+ sand 1400g is (thick：In=1:2), respectively in low-phosphorous (50 μM of KH2PO4) low (500 μM of nitrogen
Ca(NO3)2·4H2O), it is not inoculated under the conditions of (- Ri) and Rhizobium Inoculation (+Ri) after handling 6 weeks and harvests, low-phosphorous low nitrogen processing
K+Use K2SO4Polishing.As a result it shows：It is low-phosphorous connect rhizobium under the conditions of, overexpression GmSUT6 genes significantly reduce soybean
Root nodule fresh weight shows that when overexpression GmSUT6 genes, plant excessively exports photosynthate and gives root nodule homobium, root nodule life
Length is similarly subjected to inhibit.GmSUT6 has regulated and controled the output of photosynthate and the growth of root nodule.
5, the influence of overexpression GmSUT6 gene pairs Soybean transgenic plant biomass：
Biomass measures：1 percent balances weigh overground part and root samples fresh weight, all samples are dried at 105 DEG C
Case water-removing is placed on 75 DEG C in 30 minutes, and drying to constant weight, weighs dry weight.
Fig. 6 B are the influence of overexpression GmSUT6 gene pairs Soybean transgenic plant biomass.Wherein, plant uses sand
Earth culture is tested, sand ratio：Native 600g+ sand 1400g is (thick：In=1:2), respectively in low-phosphorous (50 μM of KH2PO4) low (500 μM of nitrogen
Ca(NO3)2·4H2O), it is not inoculated under the conditions of (- Ri) and Rhizobium Inoculation (+Ri) after handling 6 weeks and harvests, low-phosphorous low nitrogen processing
K+Use K2SO4Polishing.As a result it shows：It is low-phosphorous connect rhizobium under the conditions of, overexpression GmSUT6 genes significantly reduce soybean
Biomass shows that when overexpression GmSUT6 genes, excessively export photosynthate gives root nodule homobium to plant, can inhibit certainly
Body is grown.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, it is other it is any without departing from the spirit and principles of the present invention made by changes, modifications, substitutions, combinations, simplifications,
Equivalent substitute mode is should be, is included within the scope of the present invention.
Sequence table
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<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT3 F
<400> 7
tggtcccaca gataatagtg tc 22
<210> 8
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT3 R
<400> 8
gccaacacag ctatgagtcc 20
<210> 9
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT4 F
<400> 9
caaatatcct aatggctgtc tgct 24
<210> 10
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT4 R
<400> 10
cgctgtaagt gattgcaagt g 21
<210> 11
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT5 F
<400> 11
ctctcacttg gttgtcttgg 20
<210> 12
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT5 R
<400> 12
aaatgcacct tctctaactc c 21
<210> 13
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT6 F
<400> 13
ctaagatggc ccaacattct c 21
<210> 14
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT6 R
<400> 14
gctgtaagta atcgcgagtg 20
<210> 15
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT7 F
<400> 15
ctcttgctat tactttcagc gt 22
<210> 16
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT7 R
<400> 16
caccaccaaa ccacttatcc 20
<210> 17
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT8 F
<400> 17
ctctcacaac aatggaggag c 21
<210> 18
<211> 23
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT8 R
<400> 18
acatagccaa ataaatgagg ccc 23
<210> 19
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT9 F
<400> 19
caaggtctat ctttgggtgt c 21
<210> 20
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT9 R
<400> 20
gaaagtgaaa catgtaaagc cc 22
<210> 21
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT10 F
<400> 21
ttcctcttgc gattactttc ag 22
<210> 22
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT10 R
<400> 22
aaacaaagca tcccaaggac 20
<210> 23
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT11 F
<400> 23
tccaatagta cgcaaactga g 21
<210> 24
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> GmSUT11 R
<400> 24
ccatgacaac caagtgagag 20
<210> 25
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 25
atcgcccggg cctgctgcta caatatggag 30
<210> 26
<211> 31
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 26
atcgctcgag cacggtcaac tctgattagt g 31
<210> 27
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 27
atcggaattc agcgtataag catactaaac 30
<210> 28
<211> 31
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 28
atcgtctaga attgtagcag caggaagaaa g 31
<210> 29
<211> 51
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 29
ggggacaagt ttgtacaaaa aagcaggctt catggagcct ctctcttcca c 51
<210> 30
<211> 51
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 30
ggggaccact ttgtacaaga aagctgggtc cacggtcaac tctgattagt g 51
<210> 31
<211> 32
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 31
atcgagatct gacctgctgc tacaatatgg ag 32
<210> 32
<211> 31
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 32
atcgacgcgt cacggtcaac tctgattagt g 31
<210> 33
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 33
gctgttatgc ggccattgtc 20
<210> 34
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 34
gacgtctgtc gagaagtttc 20
Claims (7)
1. applications of the soybean sucrose transporter important gene GmSUT6 in low-phosphorous root nodule in terms of induced expression.
2. application according to claim 1, it is characterised in that：GmSUT6 is in plant for soybean sucrose transporter important gene
Application in breeding.
3. application according to claim 1, it is characterised in that：Soybean sucrose transporter important gene GmSUT6 is being cultivated
Application in genetically modified plants.
4. application according to claim 1, it is characterised in that：Soybean sucrose transporter important gene GmSUT6 is regulating and controlling
Plant and the application in rhizobium symbiosis.
5. application according to claim 1, it is characterised in that：It is prepared by soybean sucrose transporter important gene GmSUT6
Regulate and control plant and the application in rhizobium symbiosis preparation.
6. according to claim 2~4 any one of them application, it is characterised in that：
The plant is dicotyledonous legume.
7. application according to claim 6, it is characterised in that：
The dicotyledonous legume is soybean.
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| CN201810441185.8A CN108467869A (en) | 2018-05-10 | 2018-05-10 | The application of soybean sucrose transporter important gene GmSUT6 |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810441185.8A CN108467869A (en) | 2018-05-10 | 2018-05-10 | The application of soybean sucrose transporter important gene GmSUT6 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001088139A2 (en) * | 2000-05-12 | 2001-11-22 | Washington State University Research Foundation | Proteins interacting with a sucrose transporter |
| CN102250227A (en) * | 2010-05-17 | 2011-11-23 | 中国科学院植物研究所 | Plant sucrose transport protein, and coding gene and application thereof |
| CN102757969A (en) * | 2012-06-21 | 2012-10-31 | 华南农业大学 | Phosphorus transportprotein gene GmPT5 related to phosphorus transport of soybean nodulation and application thereof |
| CN105821060A (en) * | 2016-05-03 | 2016-08-03 | 河南农业大学 | Soybean low-phosphorus-resistance related gene GmACP2, encoded protein and application of GmACP2 |
-
2018
- 2018-05-10 CN CN201810441185.8A patent/CN108467869A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001088139A2 (en) * | 2000-05-12 | 2001-11-22 | Washington State University Research Foundation | Proteins interacting with a sucrose transporter |
| CN102250227A (en) * | 2010-05-17 | 2011-11-23 | 中国科学院植物研究所 | Plant sucrose transport protein, and coding gene and application thereof |
| CN102757969A (en) * | 2012-06-21 | 2012-10-31 | 华南农业大学 | Phosphorus transportprotein gene GmPT5 related to phosphorus transport of soybean nodulation and application thereof |
| CN105821060A (en) * | 2016-05-03 | 2016-08-03 | 河南农业大学 | Soybean low-phosphorus-resistance related gene GmACP2, encoded protein and application of GmACP2 |
Non-Patent Citations (2)
| Title |
|---|
| ALDAPE MJ等: "Identification and characterization of a sucrose transporter isolated from the developing cotyledons of soybean", 《ARCH BIOCHEM BIOPHYS.》 * |
| 张利: "杨树糖转运体SWEETs和SUT4的功能分析", 《中国博士学位论文全文数据库》 * |
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